I have frozen DNA samples and I want to do PCR on them. I want to know what I have to do with the DNA samples after getting them out of the freezer to ensure the DNA is completely soluble in the TE solution.
It would help to know what the DNA is frozen in. If it is water or TE buffer, no homogenisation is necessary. If it is lyophilised in some way, resuspending it in water should be suffcient.
Hi,
you just have to resuspend them in TE, or, better ddH20, that makes them more suitable for a PCR application.
If those samples are ethanol purified, leave the vials open in a chemical hood, and when the ethanol is gone, add water.
First, you may better boil the sample at optimal temperature.
Second, solublution is important for you
you may use molecular biology by yourself
As per your query you have frozen DNA sample, that means the DNA is dissolved in either 1X TE or pyrogen free water and stored at -20oC or at lower temperature after you isolated and purified the DNA. Am I right? If this is the case you just need to take out the frozen sample and thaw and make the necessary dilution (required for your PCR) using PF water. Hope this will help since DNA is readily soluble in 1X TE as well as PF water.
1. Thaw the frozen DNA, invert the tube gently several times, take the amount of DNA you need for PCR.
2. Aliquot your DNA stock in several tubes for storage. Avoid store all your DNA stock in one single tube and go through freeze-and-thaw cycles to get your DNA for different experiments. Repeat freeze-and-thaw cycles can reduce your DNA quality.
Dear Noha,
A good DNA prep, free of contaminating proteins, is always soluable 100% in ddwater or IXTE buffer. However, have the habit of aliquoting your DNA prep the first time around in i.e. 10 microlitter each for down stream manipulation.
Regards,
GCK
Nothing special. Just thaw them, then tap a couple of times on the tube to homogenise the solution and finally spin them for a short time.
Thaw well and spin, then you can load 5 ul in agarose gel (1%) to see bands
TE is the best for dissolving your DNA sample as well as for longer storage. After adding your TE buffer into your DNA sample I would recommend you to go for hydration step i.e. keep that vial at 4 C for at least 30 min. these will help for better DNA dissolution without affecting the quality.
Add TE to yoru DNA samples and keep on desk for 30 mins. Use pipette to dissolve.
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